[Truncated abstract] The analysis and subsequent interpretation of trace evidence is of paramount importance to the forensic scientist. While a variety of methods are available to facilitate comparison between recovered and control samples, the use of a specific analytical method depends upon both the physical and chemical nature of the material itself and the material to which it is to be compared. Elemental analysis of evidentiary material is one such method of sample comparison and has been extensively applied to this purpose following the introduction of neutron activation analysis in the early 1960s. However, over the last 15 years, another instrumental technique has taken centre stage in the analytical armoury of the forensic scientist: laser ablation–inductively coupled plasma-mass spectrometry (LA-ICP-MS). The modification and adaptation of this technique, to a point where it is possible to distinguish between glass materials produced only hours apart on the same production line, is detailed in this thesis. Additional protocols have also been developed for the analysis of fibreglass and plastic crime scene debris. Finally, a method for quantification of elemental concentrations in headlamp plastics has also been developed to facilitate inter-comparison of data between both different analytical techniques and different laboratories. Glass material is one of the most common varieties of trace evidence and the forensic examination of glass traditionally involves the determination of its refractive index (RI). ... The analytical protocol involves the analysis of 46 analytes on material comprising the exterior surface of the lens. Using this data, it was found that although minor variations in elemental composition exist within a single headlamp lens, discrimination between lenses produced from a single manufacturing plant over a short period of time could still be achieved. Discrimination between all headlamp lenses, with the exception of some lenses produced on the same day, could be facilitated using the analytical protocol developed. Furthermore, an interpretational protocol has been developed that has successfully classified all unknown headlamp lens samples investigated in this study, within the discrimination limits of the analytical method. The semi-quantitative analysis of glass and plastic samples has also been examined using LA-ICP-MS. The concentrations of 16 analytes in container and float glass samples were determined. However, the levels of discrimination afforded by the semi-quantitative data were inferior to those achieved using qualitative data. Finally, a series of plastic-based standards, containing 25 analytes of known concentrations, was produced. Using these standards, relative concentrations of the study analytes were determined in polycarbonate headlamp lenses. Interpretation of the data produced made it possible to discriminate between all study samples. Consequently, the total analytical and interpretational protocol developed in this study has established the foundation for LA-ICP-MS to be adopted internationally as a recognised method for the analysis of plastic crime scene debris.

[Truncated abstract] The sale of fraudulent South-East Asian ceramics constitutes a large proportion of the illegal artefact and antique trade and threatens to undermine the legitimate international market. The sophistication and skill of forgers has reached a level where, using traditional appraisal by eye and hand, even the most experienced specialist is often unable to distinguish between a genuine and fraudulent piece. In addition, the current scientific method of authentication used by the international antiques and art industry, thermoluminescence (TL) dating, carries severe limitations. The technique has an error margin of +/-20 % and requires the removal of a significant piece of the sample being tested, which decreases both the monetary and cultural value of the artefact. Of more concern, forgers have developed methods which produce false test results and which appear to corroborate false claims for the age of artefacts. Consequently, the use of TL dating for authentication of ancient ceramics, especially those of South-East Asian origin, has now come into serious question. The most suitable method for authenticating ceramics is through provenance establishment. Studies published in the literature have investigated the application of various analytical techniques to provide this information for ceramic wares and have highlighted their potential to be used for provenance establishment. However, the value of each of these techniques is limited rendering them generally unsuitable for practical use in the international antiques and art world to authenticate high-value South-East Asian artefacts. Consequently, there is a desperate need for the development of a robust, accurate and non-destructive method which can be practically applied in the industry to authenticate South-East Asian ceramics. ... Minor variations between spectral profiles of artefacts produced in the same country have also been used to further provenance artefacts to a specific production region or kiln site. The results of analyses have been compiled to form a unique reference database which can be added to in the future and used by experts internationally. Adaptation of the developed sampling and analytical methodologies to allow in-situ sampling of large artefacts using the

Indiana University-Purdue University Indianapolis (IUPUI) / Automotive paints have used an ultraviolet (UV) absorbing clear coat system for nearly thirty years. These clear coats have become of forensic interest when comparing paint transfers and paint samples from suspect vehicles. Clear coat samples and their ultraviolet absorbers are not typically examined or characterized using Raman spectroscopy or microspectrophotometry (MSP), however some past research has been done using MSP. Chemometric methods are also not typically used for this characterization. In this study, Raman and MSP spectra were collected from the clear coats of 245 American and Australian automobiles. Chemometric analysis was subsequently performed on the measurements. Sample preparation was simple and involved peeling the clear coat layer and placing the peel on a foil-covered microscope slide for Raman or a quartz slide with no cover slip for MSP. Agglomerative hierarchical clustering suggested three classes of spectra, and principal component analysis confirmed this. Factor loadings for the Raman data illustrated that much of the variance between spectra came from specific regions (400 – 465 cm-1, 600 – 660 cm-1, 820 – 885 cm-1, 950 – 1050 cm-1, 1740 – 1780 cm-1, and 1865 – 1900 cm-1). For MSP, the regions of highest variance were between 230 – 270 nm and 290 – 370 nm. Discriminant analysis showed that the three classes were well-differentiated with a cross-validation accuracy of 92.92% for Raman and 91.98% for MSP. Analysis of variance attributed differentiability of the classes to the regions between 400 – 430 cm-1, 615 – 640 cm-1, 825 – 880 cm-1, 1760 – 1780 cm-1, and 1860 – 1900 cm-1 for Raman spectroscopy. For MSP, these regions were between 240 – 285 nm and 300 – 370 nm. External validation results were poor due to excessively noisy spectra, with a prediction accuracy of 51.72% for Raman and 50.00% for MSP. No correlation was found between the make, model, and year of the vehicles using either method of analysis.

Drug facilitated assault (DFA) is an increasing problem in the UK. The crime often occurs through the surreptitious administration of a drug into a victims drink, rendering the victim unable to resist the assault. The detection of these drugs in a biological specimen from the victim is one of the most challenging facets of forensic chemistry. Drug concentrations can be very low, as often only a single dose is administered, and the pharmacodynamics of commonly employed drugs further hinders the testing process. The research presented in this work shows the development of several new assays for the detection of flunitrazepam, gamma-hydroxybutyrate (GHB) and ethyl glucuronide (EtG) in a variety of biological matrices. New methods of drug testing in blood and urine are demonstrated, as well as interesting developments in the field of hair testing. Using hair to detect drug exposure allows a much wider window of detection than the more traditional matrices of blood and urine. New methods are presented in this work using gas chromatography-tandem mass spectrometry (GCMS/MS) to detect drugs in hair. Validation data is presented along with the results of authentic DFA testing. All aspects of the drug testing procedure have been evaluated, from new extraction techniques utilising water instead of solvents, to novel clean up stages involving the unique combination of SFE and SPME. Several confirmation techniques are explored including single quadrupole, triple quadrupole and ion trap mass spectrometry. In addition to developing assays for DFA cases, the versatility of this type of analytical chemistry is explored in two population studies. The first study evaluates alcohol consumption between two groups; drugs users and non drug users in medico-legal cases. There is an anecdotal belief amongst drug clinic staff that alcohol use is lower in drugs users than it is in non drug users. This study presents the first scientific confirmation of this belief through EtG (an alcohol metabolite) testing in hair of the two groups. The second study investigates whether there is a correlation between EtG and cocaethylene (a metabolite of cocaine only produced in the presence of alcohol) in cocaine users. Results f this study suggest that there is no positive correlation between the two compounds. The research presented in this thesis aims to further the analytical science surrounding FA investigation and provide accurate, sensitive and reliable methodology for drug esting in blood, urine and hair.

Indiana University-Purdue University Indianapolis (IUPUI) / Toner analysis has become an area of increased interest due to the wide availability of laser printers and photocopiers. Toner is most often encountered on paper in questioned document analysis. Because of this, it is important to develop methods that limit the interference of paper without damaging or destroying the document. Previous research using Fourier transform infrared spectroscopy (FTIR) has differentiated toners based on their polymer resin components. However, Raman spectroscopy and chemometric analysis are not typically used for the examination of this material. Raman spectroscopy is a popular tool for the chemical analysis of pigmented samples and was used to characterize cyan, yellow, and magenta toners. Analyses were performed using a dispersive micro-Raman spectrometer equipped with a 785nm diode laser, a CCD detector, and an objective at 20X magnification. One hundred samples of each color toner were collected. Three different and separate methods were developed for cyan, yellow, and magenta toners on paper to optimize results. Further analysis of the magenta toners was excluded due to a weak signal and significant paper interference. The data collected from the analyses of the blue and yellow toners was then processed using a combination of statistical procedures, including principal component analysis (PCA), agglomerative hierarchal clustering (AHC), and discriminative analysis (DA). Ninety-six blue toners were analyzed by PCA and three classes of spectra were suggested. Discriminant analysis showed that the three classes were well-differentiated with a cross-validation accuracy of 100% for the training set and 100% cross-validation accuracy for the external validation set. Eighty-eight yellow toners were analyzed by AHC and four classes of spectra were suggested. Discriminant analysis showed good differentiation between the classes with a cross-validation accuracy of 95.45% for the training set, but showed poor differentiation for the external validation set with a cross-validation accuracy of 72%. While these toners were able to be discriminated, no correlation could be made between the manufacturer, printer make and model, and the toner sample.

Indiana University-Purdue University Indianapolis (IUPUI) / Forensic DNA profiling is widely used as an identification tool for associating an individual with evidence of a crime. Analysis of a DNA sample involves observation of data in the form of an electropherogram, and subsequently annotating a DNA “profile” from an individual or from the evidence. The profile obtained from the evidence can be compared to reference profiles deposited in a national DNA database, which may include the potential contributor. Following a match, a random match probability is calculated to determine how common that genotype is in the population. This is the probability of obtaining that same DNA profile by sampling from a pool of unrelated individuals. Each state has adopted various laws requiring suspects and/or offenders to submit a DNA sample for the national database (such as California’s law that all who are arrested must provide a DNA sample). These profiles can then be associated with past unsolved crimes, and remain in the database to be searched in the event of future crimes. In the case of database samples, a physical sample of the offender’s DNA must be kept on file in the laboratory indefinitely so that in the event of a database hit, the sample is able to be retested. Current methods are to collect a buccal swab or blood sample, and store the DNA extracts under strict preservation conditions, i.e. cold storage, typically -20° C. With continually increasing number of samples submitted, a burden is placed on crime labs to store these DNA extracts. A solution was required to help control the costs of properly storing the samples. FTA™ paper was created to fulfill the need for inexpensive, low maintenance, long term storage of biological samples, which makes it ideal for use with convicted offender DNA samples. FTA™ paper is a commercially produced, chemically treated paper that allows DNA to be stored at room temperature for years with no costly storage facilities or conditions. Once a sample is required for DNA testing, a small disc is removed and is to be used directly in a PCR reaction. A high quality profile is important for comparing suspect profiles to unknown or database profiles. A single difference between a suspect and evidentiary sample can lead to exclusion. Unfortunately, the DNA profile results yielded from the direct addition have been unfavorable. Thus, most crime laboratories will extract the DNA from the disc, leading to additional time and cost to analyze a reference sample. Many of the profiles from the direct addition of an FTA™ disc result in poor quality profiles, likely due to an increase in PCR inhibitors and high concentrations of DNA. Currently, standardized protocols regarding the recommended locations for removal of a sample disc from a bloodspot on an FTA™ card does not exist. This study aims to validate the optimal location by comparing DNA profiles obtained from discs removed from the center, halfway, and edge locations of a bloodspot from 50 anonymous donors. Optimal punch location was first scored on the number of failed, partial or discordant profiles. Then, profile quality was determined based on peak characteristics of the resulting DNA profiles. The results for all three disc locations were 5.3% failed amplifications, 4.2% partial amplifications, and one case of a discordant profile. Profile quality for the majority of the samples showed a high incidence of stutter and the absence of non-template adenylation. Of the three disc locations, the edge of the blood stain was ideal, due to a presumably lower concentration of DNA and likely more dilute amount of the PCR inhibitor heme. Therefore, based on the results of this study, there is a greater probability of success using a sample from the edge of a blood stain spotted in FTA™ paper than any other location of the FTA™ card.

Indiana University-Purdue University Indianapolis (IUPUI) / Organic-rich substrates such as soil provide an excellent carbon source for bacteria. However, hydrocarbons such as those found in various ignitable liquids can also serve as a source of carbon to support bacterial growth. This is problematic for fire debris analysis as samples may be stored at room temperature for extended periods before they are analyzed due to case backlog. As a result, selective loss of key components due to bacterial metabolism can make identifying and classifying ignitable liquid residues by their chemical composition and boiling point range very difficult. The ultimate goal of this project is to preserve ignitable liquid residues against microbial degradation as efficiently and quickly as possible. Field and laboratory studies were conducted to monitor microbial degradation of gasoline and other ignitable liquids in soil samples. In addition to monitoring degradation in potting soil, as a worst case scenario, the effect of soil type and season were also studied. The effect of microbial action was also compared to the effect of weathering by evaporation (under nitrogen in the laboratory and by the passive headspace analysis of the glass fragments from the incendiary devices in the field studies). All studies showed that microbial degradation resulted in the significant loss of n-alkanes and lesser substituted alkylbenzenes predominantly and quickly, while more highly substituted alkanes and aromatics were not significantly affected. Additionally, the residential soil during the fall season showed the most significant loss of these compounds over the course of 30 days. To combat this problem, a chemical solution is to be immediately applied to the samples as they are collected. Various household and commercial products were tested for their efficacy at low concentrations to eliminate all living bacteria in the soil. Triclosan (2% (w/v) in NaOH) proved to be the most effective at preserving ignitable liquid residues for at least 30 days.

Indiana University-Purdue University Indianapolis (IUPUI) / Clear coats have been part of automotive field paint finishes for several decades. Originally a layer of paint with no pigment, they have evolved into a protective layer important to the appearance and longevity of the vehicle's finish. These clear coats have been studied previously using infrared spectroscopy and other spectroscopic techniques. Previous studies focused on either all the layers of an automobile finish or on chemometric analysis of clear coats using other analytical techniques. For this study, chemometric analysis was performed on preprocessed spectra averaged from five separate samples. Samples were analyzed on a Thermo-Nicolet Nexus 670 connected to a Continuμm™ FT-IR microscope. Two unsupervised chemometric techniques, Agglomerative Hierarchical Clustering (AHC) and Principal Component Analysis (PCA), were used to evaluate the data set. Discriminant analysis, a supervised technique, was evaluated using several known qualifiers; these included cluster group from AHC, make, model, and year. Although discriminant analysis confirmed the AHC and PCA results, no correlation to make, model, or year was indicated.